Use of alternan as texturizing agent in foodstuffs and cosmetics

ABSTRACT

The invention is directed to the use of alternan as texturizing agent, particularly as fat or oil replacer in foodstuffs or cosmetic preparations, a homogeneous composition comprising alternan and water, the use of the homogeneous composition as texturizing agent in foodstuffs or cosmetic preparations, and foodstuffs and cosmetic preparations comprising alternan as texturizing agent or a homogeneous composition comprising alternan.

CROSS REFERENCE TO RELATED APPLICATION

This application is a Divisional application Ser. No. 12/921,046 filedNov. 18, 2010, which claims priority to U.S.C. 371 National Phase of PCTApplication No. PCT/EP2009/001760 filed Mar. 6, 2009, the disclosure ofthese prior applications are hereby incorporated in their entirety byreference.

The present invention is directed to the use of alternan as texturizingagent, particularly as fat or oil replacer in foodstuffs or cosmeticpreparations, a homogeneous composition comprising alternan and water,the use of the homogeneous composition as texturizing agent infoodstuffs or cosmetic preparations, and foodstuffs and cosmeticpreparations comprising alternan as texturizing agent or a homogeneouscomposition comprising alternan.

In recent years, problems of obesity and hyperlipemia have increased dueto excessive consumption of oils, fats, sugars, etc., and decreasedconsumption of dietary fibers. It has been suggested that certain adultdiseases, which are leading causes of death, such as cancer, heartdiseases, etc., are associated with excessive consumption of oils, fats,and sugars and decreased consumption of dietary fibers. For thesereasons, low calorie foodstuffs wherein oil or fat is replaced bysubstances with beneficial texturizing properties have become popular.

Some Polysaccharides have been described as texturizing agents and fatreplacers. U.S. Pat. No. 5,169,671 relates to a food or drink containinga fructose polymer mainly composed of beta-2,1-bond, referred to as“polyfructan”, as a substitute for oils, fats, and sugars and havingimproved gel properties compared with foods containing other oil, fat,or sugar substitutes and a creamy taste and texture equivalent to thatderived from oils and fats and a method of producing such a food ordrink.

WO2007/128559 and WO2007/128559 teach that very long chain inulins areefficient texturizing agents and fat replacers, especially in dairyproducts and ice cream.

However, in view of the increased need for efficient texturizing agentsand fat replacers for foods it was an object of the present invention tofind alternative substances which can be used for this purpose

The present invention is directed to the use of alternan as atexturizing agent in foodstuffs or cosmetic preparations.

In another aspect the present invention is directed to the use ofalteman as an emulsifier, preferably for foodstuffs and cosmeticpreparations.

A texturizing agent according to the present invention imparts textureto foodstuffs or cosmetic preparations or modifies the texture offoodstuffs or cosmetic preparations. The term “texture” refers to theproperties held and sensations caused by the external surface offoodstuffs or cosmetic preparations received through the sense of touch.

In connection with foodstuffs, the term “mouthfeel” is usedalternatively to the term “texture” and both terms are used hereininterchangeably with respect to foodstuffs. In connection with cosmeticpreparations, a major meaning of the term texture is the skin feel of acosmetic preparation.

In the present invention it can be shown that alternan is a verysuitable fat or oil replacer in foodstuffs. The fat or oil component offoodstuffs has usually a major impact on its texture/mouthfeel,organoleptic characteristics, and flavor. A fat or oil content usuallyimparts a creamy and smooth mouthfeel to a foodstuff. These desiredproperties of foodstuffs can also be reached by adding alteman tofoodstuffs. The term “fat or oil replacer” in this connection means thatat least a part of a fat or oil component in a foodstuff is removed fromthe foodstuff and replaced by alternan. The term “fat or oil replacer”,however, can also mean that the total fat or oil component in afoodstuff is removed from the foodstuff and replaced by alternan.

Alternan is a polysaccharide composed of glucose units. The glucoseunits are linked to each other via a-1,3- and a-1,6-glycosidic bonds,and said two types of bonds predominantly appear alternatingly. Alternanmay contain branches (Seymour et al., Carbohydrate Research 74, (1979),41-62). Alternan and methods for producing alternan are well known fromthe state of the art. See for example Jeanes et al. (1954) J. Am. Chem.Soc., 76: 5041-5052, Misaki et al. (1980) Carbohydr. Res., 84: 273-285,Cote and Robyt (1982), Carbohydr. Res., 101: 57-74, Cote (1992),Carbohydrate Polymers 19, 249-252, WO 00/47727, U.S. Pat. No. 5,702,942,US20060127328, PCT/EP2008/051760.

Alternan according to the present invention preferably has a weightaverage molecular weight Mw in the range of 10 000 000 g/mol to 60 000000 g/mol (determined with GPC MALLS), more preferably in the range of12 000 000 g/mol to 50 000 000 g/mol. In a special embodiment, altemanis produced with alternansucrase originating from LeuconostocMesenteroides as described in WO 00/47727 and shows a weight averagemolecular weight Mw in the range of 33 000 000 g/mol to 60 000 000 g/mol(determined with GPC MALLS), more preferably in the range of 33 000 000g/mol to 50 000 000 g/mol. In still another special embodiment, alternanis produced with truncated alternansucrase enzyme as described inPCT/EP2008/051760 and shows a weight average molecular weight Mw in therange of 12 000 000 g/mol to 30 000 000 g/mol (GPC MALLS), morepreferably in the range of 14 000 000 g/mol to 28 000 000 g/mol, stillmore preferably in the range of 16 000 000 g/mol to 26 000 000 g/mol,most preferably 19 000 000 g/mol to 23 000 000 g/mol. Truncatedalternansucrase enzymes, methods for producing alternan therefrom aswell as the alternan itself are described in PCT/EP2008/051760, which isincorporated herein by reference in its entirety.

A preferred foodstuff wherein alteman is used as a fat replacer isselected from dairy products, yoghurts, ice creams, milk-based soft ice,milk-based garnishes, puddings, cream, whipped cream, chocolate cream,butter cream, crème fraiche, curd, milk, such as skim milk, buttermilk,soured milk, kefir, milkshakes, egg custard, cheese, such as creamcheese, soft cheese, sliced cheese, hard cheese, nutrition bars, energybars, breakfast bars, confectionery, bakery products, crackers, cookies,biscuits, cereal chips, snack products, diet drinks, finished drinks,sports drinks, stamina drinks, powdered drink mixtures for dietarysupplementation, infant and baby foodstuff, bread, croissants, breakfastcereals, spreads, sugar-free biscuits and chocolates, calcium chews,meat products, sausages, mayonnaise, dressings, nut butter, deep-frozenmeals, sauces, gravy, soups, shortenings, canned foods andready-to-serve meals.

The level of alternan in the foodstuff depends on the kind of foodstuff.Usually, alteran is used in an amount which is sufficient to reach thedesired degree of fat/oil replacement and the desired mouthfeelproperties. Without limitation exemplary levels of alteman in foodstuffsare 0.1-10 weight percent, more preferably 0.1-5 weight percent, andmost preferably 0.1-3 weight percent, based on the total weight of allcomponents forming the foodstuff.

Oils and fats may be wholly substituted with alternan. However, apreferred degree of substitution is 20 to 80% by weight based on weightof the oils and/or fats.

In a further aspect the present invention is directed to a foodstuffcomprising alternan as texturizing agent, particularly as a fat or oilreplacer. The foodstuff is preferably selected from the above mentionedfoodstuffs The term “foodstuff” according to the invention alsoencompasses beverages.

The present invention is also directed to a cosmetic preparation,comprising alternan as texturizing agent. The cosmetic preparation wherealternan may be employed include, but are not limited to, deodorants,antiperspirants, antiperspirant/deodorants, shaving products, skincreams and lotions, moisturizers, toners, bath products, cleansingproducts, hair care products such as shampoos, conditioners, mousses,styling gels, nails creams and lotions, cuticle softeners, protectivecreams such as sunscreen, anti-aging products, and other personal careformulations.

In a further aspect, the present invention is directed to a homogeneouscomposition comprising alternan and water. The term “homogeneouscomposition” means that the composition shows no visible phaseseparation or separation of its constituents (not visible to the nakedeye).

Depending on the relative amounts of alternan and water, saidhomogeneous composition can show a wide viscosity range. So, thehomogeneous composition may be a liquid or a more viscous cream, whereina cream is preferred.

In one embodiment the composition according to the invention comprises5-15 weight-% alternan and 85-95 weight-% water, based on the totalweight of the composition, more preferably 8-13 weight-% alternan and87-92 weight-% water. Further components may be included, such assurfactants and salts.

In another embodiment, a homogeneous composition according to theinvention comprises alternan, water and at least one fat or oil or amixture of fat(s) and oil(s). Said homogeneous composition comprisingalternan, water and at least one fat and/or oil shows no visible phaseseparation between the fat or oil component and the water component (notvisible to the naked eye). Thus, said composition comprising alternan,water and at least one fat and/or oil may also be designated as anemulsion.

Depending on the relative amounts of alternan, water, and oil and/orfat, said homogeneous composition can show a wide viscosity range. So,the homogeneous composition may be a liquid or a more viscous cream,wherein a cream is preferred.

In one embodiment a composition comprises 5-15 weight-% alternan, 70-90weight-% water, 5-15 weight-% fat or oil, more preferably 8-13 weight-%alternan, 74-84 weight-% water, and 8-13 weight-% fat or oil, based onthe total weight of the composition. Further components such assurfactants and salts may be included.

Preferred fats/oils as a component for a homogeneous compositioncomprising alternan, water and at least one fat and/or oil are selectedfrom arachidic oil, avocado oil, cotton seed oil, safflower oil, peanutoil, hazelnut oil, hemp oil, jojobaoil, camenillia oil, cocoa butter,coconut oil, pumpkin oil, linseed oil, macadamia nut oil, corn germ oil,almond oil, mango seed fat, poppy seed oil, evening primrose oil, oliveoil, palm oil, palm kernel oil, papaya oil, pistachio nut oil, pecan nutoil, rapeseed oil, castor oil, mustard seed oil, sesame oil, rheabutter, soybean oil, sunflower oil, walnut oil, water melonseed oil,grapeseed oil, wheat germ oil, cedarwood oil, wherein said oils mightoptionally be hardened oils.

In still another aspect, the present invention is directed to ahomogeneous composition comprising alternan and milk. In a preferredembodiment said composition comprises 5-15 weight-% alternan and 85-95weight-% milk, based on the total weight of the composition, morepreferably 8-13 weight-% alternan and 87-92 weight-% milk. Furthercomponents may be included, such as surfactants and salts. A creamcomprising alternan and milk might preferably be used for all kind ofdairy products.

The viscosity of a homogeneous composition according to the invention isadapted to the desired kind of foodstuff or cosmetic preparation, i.e.to the desired final viscosity of said foodstuff or cosmetic. However,as said above, the homogeneous composition is preferably a cream,wherein a preferred range of viscosity of a cream is from 0.5 to 1.5Pa*s, more preferably 0.8 to 1.3 Pa*s at 20° C. and a shear rate of 40s⁻¹.

All of the above described homogeneous compositions, preferably creams,are hereinafter also designated as “(homogeneous) composition accordingto the invention” or “cream according to the invention”.

The present invention also relates to a method for preparing the abovedescribed compositions, the method comprising

-   -   a) giving alternan and water into a vessel    -   b) applying shear to obtain a homogeneous composition, and        optionally    -   c) adding oil or fat and applying shear to obtain a homogeneous        composition.

The optional step c) relates to the above described embodiment of ahomogeneous composition, comprising alteman, water and oil or fat.

Mixing in steps b) and c) is preferably done under high shear withcommonly known devices for such purpose, such as ULTRA TURRAX®dispersing tool from IKA company. In a preferred embodiment, steps a)and b) are performed simultaneously, e.g. by adding alteman as a powderto water with simultaneous application of high shear. Optionalcomponents as salts and emulsifiers might be added at any stage of thepreparation.

In a further aspect, the present invention is related to the use of ahomogeneous composition as described above for the manufacture offoodstuffs or cosmetic preparations. In this connection, saidhomogeneous compositions, preferably creams, can preferably used forabove-listed foodstuffs and cosmetics.

According to the present invention the homogeneous compositions can beused as constituent for foodstuffs or cosmetic preparations. In thisconnection, homogeneous compositions according to the present inventioncan fulfill, without limitation, following functions: giving body tofoodstuffs or cosmetic preparations, act as a base for foodstuffs orcosmetic preparations, act as a texturizing agent for foodstuffs orcosmetic preparations. The homogeneous compositions according to thepresent invention can fulfill one or more of the aforementionedfunctions.

In regard to texturizing properties the homogeneous compositionsaccording to the present invention are preferably used as a fat or oilreplacer in foodstuffs. In this connection, the homogeneous compositionis preferably a cream. The preferred foodstuffs for said use areselected from dairy products, yoghurts, ice creams, milk-based soft ice,milk-based garnishes, puddings, cream, whipped cream, chocolate cream,butter cream, crème fraiche, curd, milkshakes, egg custard, cheese, suchas cream cheese, soft cheese, sliced cheese, hard cheese, nutritionbars, energy bars, breakfast bars, confectionery, bakery products,crackers, cookies, biscuits, cereal chips, snack products, infant andbaby foodstuff, bread, croissants, spreads, sugar-free biscuits andchocolates, calcium chews, meat products, sausages, mayonnaise,dressings, nut butter, deep-frozen meals, sauces, gravy, soups,shortenings, canned foods and ready-to-serve meals.

Homogeneous compositions according to the invention have a texturesimilar to oils or fats. By substituting this composition for liquidoils and solid fats, for example in dressings, mayonnaise, fresh cream,cream cheese, butter, salad oil, etc., in various cooked and processedfood, the number of calories in the resulting food are greatly reduced.Oils and fats may be wholly substituted with a homogeneous compositionaccording to the present invention. However, a preferred degree ofsubstitution is 20 to 80% by weight based on the oils and fats.

Finally, the present invention is also directed to a foodstuff orcosmetic preparation, characterized in that during its manufacturingprocess a homogeneous composition according to the present invention wasadded as a constituent. In the foodstuff or cosmetic, a homogeneouscomposition according to the present invention preferably fulfills thefunctions as described above with respect to its use. Depending on thekind of foodstuff or cosmetic and its manufacturing process, thehomogeneous composition might be present in the final product withoutsubstantial structural alteration. This might for example be the casewhen a cream of the invention is used as the base for a cosmeticpreparation and only some active ingredients are added. In such case,the final foodstuff or cosmetic preparation is characterized in that itstill comprises the cream. In other cases, e.g. in products which areheat treated after addition of the cream, the cream might be subjectedto major alterations in its structure. The present invention is to beunderstood to encompass all foodstuffs and cosmetic preparations duringwhose manufacturing process a cream according to the invention wasadded, regardless of whether the initial structure of the cream isaltered in the final product or not.

In the following section, the present invention is further illustratedby examples, which are, however, not intended to limit the scope of theinvention as defined in the appended claims.

EXAMPLES

1. Manufacture of Alternan

Plasmid pAI-B-AlSu Q29 was transformed in E. coli DHSa. VectorpAI-B-AlSu contains the full-length coding sequence of alternansucrasederived from Leuconostoc mesenteroides strain NRRL B-1355 (cf. WO00/47727), lacking the N-terminal 39 amino acids from the signalpeptide, fused to an octapeptide strep-tag at the C-terminal end. Thestrep-tag is linked to the protein through a dipeptide linker.Expression of alternansucrase is under the transcriptional control ofthe tetA promoter/operator and repressor. The tetA promoter is tightlyregulated by the tet repressor which is encoded on the same plasmid andis constitutively expressed from the β-lactamase promoter. In this way,expression of alternansucrase is stringently repressed until efficientchemical induction by tetracycline or anhydrotetracycline, AHT.

The cells were pre-cultured in mineralmedium (Horn et al., 1996) with100 μg/ml Ampicillin and 10% LB medium. Mineralmedium, without LB, wasinoculated with this pre-culture. The cells were grown at 37° C.,induced with Anhydrotetracyclin (AHT) (0.2 mg/L), and grown further at25° C. The cells were harvested, resuspended in [10 mM MOPS pH 7.6; 2.5mM CaCl2 and 0.05% Trition X-100] and extracted with a high pressurehomogenisator. The cell lysate was centrifuged at 20 000 rpm for 20minutes at 4° C. The supernatant was filtered over a 0.22 μm filter.

Alternan was produced in a 60 L Biotransformation containing 0.13%Acetic Acid, 100 mM NaAc pH5.3, 20% Sucrose, 1 mM DTT, 1600 ml filteredprotein extract (ca. 3900 Units). The reaction mixture was incubated for60 h. at 37° C. The polymer was precipitated with 60 L Technical Ethanol40 h 4° C., washed 2× with 60 L 60% Technical Ethanol, and 1× with 60 L60% Ethanol Absolute. The product was dried through lyophilization

Reference:

-   Horn U, Strittmatter W, Krebber A, Knupfer U, Kujau M, Wenderoth R,    Muller K, Matzku S, Pluckthun A, Riesenberg D. High volumetric    yields of functional dimeric miniantibodies in Escherichia coli,    using an optimized expression vector and high-cell-density    fermentation under non-limited growth conditions. Appl Microbiol    Biotechnol 1996; (46): 524-532.

2. Application Tests

2.1 Mayonnaise

Background:

The objective was to test the alternan as produced in example 1(hereinafter “altenan”) as a fat replacer in a model mayonnaise typedressing system. A model mayonnaise dressing was developed with thecontrol mayonnaise at 68% oil (the standard of identity for mayonnaiseis >65% oil).

Three methods of incorporating the alternan were evaluated:

-   -   1. Making an alternan cream and adding it in the last step after        the oil.    -   2. Dry blending the alternan with dry ingredients and adding it        to the egg mixture.    -   3. Making an alternan cream and adding it to the egg mixture.

After the initial experiments using Test Method 1, it was determinedthat the level of alternan needed to be higher.

Fat replacement levels of 25% to 50% were tested.

Control Formula and Method:

Ingredients % Egg Mixture Egg Yolks 11.15 Water 9.17 Vinegar (DistilledWhite, 5% Acidity) 8.78 Sugar 1.30 Salt 0.81 Mustard Powder 0.64Mayonnaise Egg Mixture (from above) 31.85 Salad Oil (Soybean) 68.15

-   -   The egg yolks, water, vinegar (5% acidity), sugar, salt and dry        mustard were mixed together and heated slowly until just        beginning to boil.    -   The egg mixture was allowed to cool slightly and then soybean        oil was sheared into the mixture using a Silverson High Shear        mixer fitted with a Square Hole High Shear screen to form an        emulsion.    -   The mayonnaise was refrigerated overnight before measuring the        viscosity (sample temperatures ranged from 3.3 to 6.7° C.).        Viscosity was measured with a Brookfield RV at Speed 10 with        Spindle #5 or 6, the reading was recorded after 1 minute.

Test Method 1:

Ingredients Alternan Cream % Water 80.91 alternan 10.0 Salad Oil(Soybean) 9.09 Mayonnaise 50% Fat Repl. 25% Fat Repl. Egg Mixture (asdescribed in Control) 31.85 31.85 Salad Oil (Soybean) 34.15 50.15Alternan Cream 34.00 18.00 Final % of alternan in 50% Fat ReplacedMayonnaise = 3.4% Final % of alternan in 25% Fat Replaced Mayonnaise =1.8%

-   -   A cream was prepared with water, 10% of the alternan and 9.09%        Soybean Oil (using the Silverson High Shear mixer).    -   The mayonnaise was prepared as described in the Control method        but after the addition of the Soybean Oil, the alternan Cream        was sheared in.

“Blank”

-   -   A sample with a 25% reduction in oil was prepared by simply        increasing the water (no alternan cream was added).

Results and Observations, Test Method 1:

The formula and process for preparing the alternan cream worked well.The cream was white with a medium viscosity. However, the dressingsprepared with the alteman cream were much thinner than the control. Thetest product with a 25% reduction in oil had some viscosity and wassimilar to a pourable style dressing. The Test product with a 50%reduction in oil was very thin. The level of alternan may have been toolow.

Viscosity Sample pH Aw (centapoise)* Observations Control 4.09 0.96433,000 Thick, forms “peaks” Test with alternan, 4.27 0.984 5,500 Thin50% oil replacement Test with alternan, 4.24 0.978 9,500 Some thickness/25% replacement slight gel Test, Blank with 4.27 0.982 6,200 Somethickness/ 25% replacement slight gel

Test Method 2:

The alternan powder was dry blended with the sugar, salt and mustard.The Control procedure was then followed. A 50% reduction in fat wastested using 10% alternan and 5% alteman. The amount of water in theformula was adjusted to compensate for the reduction in oil and theaddition of the alternan to balance the formula to 100%.

Results and Observations, Test Method 2:

The 10% level of alternan was too high for this application. It was verydifficult to incorporate the alternan into the egg mixture and afoodstuff processor to mix the mayonnaise had to be used. The resultingproduct was more like a shortening or spread than a spoonable dressing.The 5% level worked well with the method as described and the resultingproduct was similar to Control with respect to appearance, texture andviscosity.

Viscosity Sample pH (centapoise) Observations Control (same as described4.09 33,000 Thick, forms “peaks” in Method 1) 50% Reduced Fat with 10%1,060,000*   Similar to shortening alternan or butter, fairly stiff andslightly gel like with a smooth/fatty feel. Oil droplets are very smalland uniform (microscopic examination). 50% Reduced Fat with 5% 39,700Thick, similar to alternan, Sample 1 control mayonnaise. Has a smooth,fatty feel when rubbed between the fingers. Very small and uniform oildroplets (microscopic examination). 50% Reduced Fat with 5% 4.14 22,000Thinner than the alternan, Sample 2 control but still like a mayonnaise.Very fine and uniform oil droplets (microscopic examination). *Since thesample was so thick the viscosity measurement procedure was modified:the Brookfield with Heliopath Spindle F was used at speed 5.

Test Method 3

Ingredients % Egg Mixture Egg Yolks 11.15 Water 0 Vinegar (DistilledWhite, 5% Acidity) 8.78 Alternan Cream* 48.25 Sugar 1.30 Salt 0.81Mustard Powder 0.64 *Alternan Cream Water 79.30 alternan 10.36 Salad Oil(Soybean) 10.34 Mayonnaise Egg Mixture 70.93 Salad Oil (Soybean) 29.07Final level of alternan = 5%

Alteman Cream

-   -   Add the alternan into the water slowly while shearing with the        Silverson mixer.    -   After all the alternan is incorporated, add the oil slowly until        well blended.    -   Follow the control procedure for the egg mixture but whisk the        alternan Cream in last.    -   Follow the control procedure for preparing the mayonnaise.

Results and Observations, Test Method 3:

The method worked well to prepare the alternan cream and it blended intothe egg mixture easily. The egg mixture was thick but flowable. Thefinished mayonnaise was thicker than the control.

Viscosity Sample pH (centapoise) Observations Control, 2^(nd) Sample4.16 35,700 Thick, typical mayonnaise. Oil droplets are mostly verysmall with a few larger ones (microscopic examination). 50% Reduced Fatwith 5% 4.17 68,000 Thicker than the alternan control but still like amayonnaise. Oil droplets look the same as control (microscopicexamination).

Discussion and Conclusions:

The alternan works well to replace 50% of the fat in a spoonabledressing/mayonnaise system at a 5.0% level. A lower level (3.4%) did notadd enough viscosity to be comparable to the control. At the 5.0% level,a fairly large variation in the sample viscosity was found whenproducing a replicate sample on a different date and when using analternate method. When the alternan was dry blended with other dryingredients and then added to the egg mixture, resulting viscositieswere 39,700 cps and 22,000 cps. However, when the alternan was firstmade into a cream and then incorporated into the egg mixture theresulting product viscosity was 68,000 cps.

2.2 Pourable Dressing

Background:

The objective was to test the carbohydrate as a fat replacer in a modelpourable dressing system. We initially tested a 50% reduction in fat andtested several oil types. We later tested a further reduction in fatwith one oil type. The reference material selected was Avicel (FMCBioPolymers, microcrystalline cellulose and carboxymethyl cellulose), acarbohydrate commonly used in low and no fat salad dressings.

Methods:

Control Test with alternan Reference Ingredients % % % Water 21.35 38.8542.6 Xanthan Gum 0.25 0.25 0.25 Propylene Glycol Alginate 0.15 0.15 0.15alternan 0.00 5.00 0.00 Avicel SD 3410 0.00 0.00 1.25 Salt 1.75 1.751.75 High Fructose Corn Syrup 21.0 21.0 21.0 Vinegar (120 grain) 10.510.5 10.5 Vegetable Oil 45.00 22.5 22.5

-   -   Dry blend the gums (and alternan or Avicel) and add them to the        water while shearing with the Silverson High Shear mixer.    -   Shear for 5 minutes. Add the salt, HFCS and vinegar and mix.    -   Slowly add the oil while shearing and shear for an additional        1.5 minutes after all the oil is added.

Note: a “blank” dressing was also prepared with 50% reduced oil but noalternan or avicel in order to determine the viscosity contribution ofthe xanthan and PGA gums. The Avicel was tested at 0.5, 1.0 and 1.25%but the highest level was closest in viscosity to the control (stilllower). An additional sample was also prepared as a reference in whichthe two gums were increased to levels recommended for a dressing ofapproximately 20% oil (0.40% xanthan gum and 0.25% PGA).

Results and Observations, Oil Comparison:

The following table summarizes the results for all the oils tested

Cold* Ambient** Viscosity Viscosity Observations and MicroscopicVariable (cps) (cps) pH Aroma (Ambient) Evaluation Control with 45%15,200 13,200 3.51 Oily, slightly painty, Pale yellow/beige color, noSoybean Oil, pungent vinegar. separation. Fairly uniform oil droplets,range about 20 to 30 microns. Control with 45% 14,000 11,040 3.50 Oily,slightly painty, Pale yellow/beige color, no Soybean Oil, pungentvinegar. separation. Fairly uniform oil droplets, range about 20 to 30microns. Test with 22.5% 12,880 9,200 3.54 Mild, slightly oily, slightWhiter/brighter than control, no Soybean Oil, painty, low pungent.separation. Small to medium oil droplets in the range of 2 to 20microns. Test with 22.5% 13,400 8,800 3.55 Mild, slightly oily, slightWhiter/brighter than control, no Soybean Oil, painty, low pungent.separation. Fairly uniform oil droplets, most roughly 6 to 10 microns.Test with 22.5% 13,320 9,200 3.54 Very mild, little to no Slightly morebeige versus the control, Corn Oil pungency. no separation. Small tomedium droplets in the range of 2 to 20 microns. Test with 22.5% 13,6809,600 3.54 Mild, slightly floral, very Slightly darker/more tan thancontrol, Canola Oil slight pungent. no separation. Small to medium oil(Rapeseed) droplets in the range of 2 to 20 microns. Test with 22.5%12,600 9,200 3.55 Mild, slightly nutty, very low Slightly whiter thanthe control, no Sunflower pungent. separation. Small to medium oil Oildroplets in the range of 2 to 20 microns. Test with 22.5% 14,280 9,2003.57 Fruity oil, no pungency. Greenish/yellow color compared to ExtraVirgin the control, no separation. Small to Olive Oil medium oildroplets in the range of 2 to 20 microns. Test “Blank” 3,440 2,720 3.42Oily, slightly painty, mild Slightly whiter/brighter than control, with22.5% pungency. no separation. Oil droplets not Soybean Oil uniform, inthe range of 5 to >40 microns. Reference with 7,440 6,400 3.54 Oily,slightly painty, mild Slightly whiter than the control, no 22.5%pungency. separation. Small to medium oil Soybean Oil, droplets in therange of 2 to 20 Increased microns. Xanthan and PGA Gums Reference with7,720 6,320 3.67 Moderate oil, slightly Slightly more white versus thecontrol, 22.5% painty, some pungency. no separation. Oil droplets in theSoybean Oil, range of 2 to 30 microns. 1.25% Avicel SD 3410 Control with45% 11,200 6,320 3.49 Very fishy. Some separation noted, 3-5 mm of TunaOil clear liquid on top, the color is more opaque on the bottom. Oildroplets are not uniform. Size range is about 2 to 100 microns. Testwith 22.5% 15,880 8,600 3.53 Somewhat fishy, not No separation noted buta slight oil Tuna Oil pungent. “slick” on the top. Oil droplets are moreuniform and smaller versus the control. Most are about 2 to 30 microns,some larger. Reference with 9,800 6,280 3.62 Somewhat fishy, not Noseparation noted but a slight oil 22.5% Tuna pungent. “slick” on thetop. Oil droplets are in Oil, 1.25% the range about 2 to 50 microns,Avicel SD 3410 some larger, many elongated versus round. *ColdViscosity: Measured on samples immediately after removing from therefrigerator (range 3.3 to 5.6° C.). **Ambient Viscosity: Measured onsamples at room temperature (range 20 to 20.6° C.).

Stability Study

Several of the samples were tested for stability by putting them at37.8° C. and evaluating them at 3 and 7 days. Note that the Tuna Oilsamples were not produced in time to include them in the study.

The following table summarizes the results of the stability test:

Results: Day 3 Day 7 “Initial” Viscosity Day 3 Viscosity Day 7 ViscosityEquilibrated Viscosity Equilibrated Viscosity Sample sample, not SampleDay 3 sample, not Sample Day 7 Sample shaken shaken shaken Observationsshaken, shaken Observations Control 12,960 13,480 12,080 No separation,12,880 10,520 Same as Day 3 color similar, oil droplets in the range of20 to 40 microns. Test with 8,880 12,680 9,360 No separation, 12,6809,200 About ¼″ of Soybean color similar, oil clear liquid noted Oildroplets in the on the bottom of range of 2 to 20 the jar. Oil dropletsmicrons. look the same as Day 3. Test with 9,000 12,280 9,200 Noseparation, 11,760 8,760 About ¼″ of Corn Oil color similar, oil clearliquid noted droplets in the on the bottom of range of 2 to 20 the jar.Oil droplets microns. look the same as Day 3. Test with 9,520 13,0409,680 No separation, 12,080 9,000 About ¼″ of Canola Oil color similar,oil clear liquid noted droplets in the on the bottom of range of 2 to 20the jar. Oil droplets microns. look the same as Day 3. Test with 9,32013,080 9,400 No separation, 12,160 9,200 About ¼″ of Sunflower colorsimilar, oil clear liquid noted Oil droplets in the on the bottom ofrange of 2 to 20 the jar. Oil droplets microns. look the same as Day 3.Test with 9,480 12,800 9,920 No separation, 13,000 9,440 About ¼″ ofOlive Oil color slightly clear liquid noted browner, oil on the bottomof droplets in the the jar. Oil droplets range of 2 to 20 look the sameas microns. Day 3. Reference 6,560 7,080 6,000 No separation, 6,4805,600 Same as Day 3 with color similar, oil (no separation Increaseddroplets in the noted). Gums range of 2 to 20 microns. Reference 6,6008,480 6,800 No separation, 8,200 6,800 No separation with 1.25% colorsimilar, oil noted. Most oil Avicel droplets in the droplets in therange of 2 to 20 range of 2 to 20 microns. can see microns, some up“fibers” under to 40 microns. microscope.

Further Reduced Fat Dressing

In order to determine if the alternan would work in further reducing thefat in the salad dressing, samples with 10% fat (reduced from 45%) wereprepared. At 10% fat, the salad dressing would qualify for a low fatclaim which is ≤3.0 grams of fat per serving, serving size is 30 grams.Two higher levels of alternan were tested: 6.5% and 7.75% (7.75% wastested first and the sample was thicker than the control). The sampleswere prepared as described previously. After preparation, the sampleswere refrigerated overnight and measured for viscosity. The samples werethen allowed to come to ambient temperature and viscosity was checkedagain. We then put the samples at 37.8° C. to test stability andevaluated at 3 and 7 days

A table summarizing the results follows:

Day 3 Day 3 Day 7 Day 7 Sample Refrig. Ambient Viscosity ViscosityViscosity Viscosity Description pH Viscosity Viscosity EquilibratedShaken Equilibrated Shaken Observations 10% SBO, 3.58 32,320 26,40033,000 33,300 41,500 38,200 No separation at 7.75% 3 days, a slightalternan “crack” near top at 7 days. Oil droplets in the range of 2 to20 microns, most are <10 microns. 10% SBO, 3.58 10,520 8,120 12,8008,640 13,080 10,200 No separation at 6.5% 3 days, a slight alternanamount of clear liquid at the bottom of the jar (about 0.25 cm) at 7days. Oil droplets in the range of 2 to 20 microns, most are <10microns.

Discussion:

The alternan works well to partially replace the fat in a reduced fatsalad dressing. The alternan acts to build back the viscosity which islost when the oil is decreased. The appearance and aroma of the testdressings was fairly comparable to the Control and the alternan may actto mask some of the pungent aroma. There didn't seem to be aninteraction between the alternan and the oil type. The thickness of thedressing seemed to be related to the iodine value of the oil used, thelower the iodine value, the less double bonds, the thicker it would tendto be at refrigerated conditions. There did seem to be a slightadvantage to using the alternan in a fish oil containing dressing. Thecontrol dressing separated and the oil droplets were larger versus thetest dressing with alternan which did not separate and had smaller oildroplets. The alteman containing dressings were able to stand up to someabuse (3 days at 37.8° C.) some separation after 7 days at 37.8° C. wasseen. Even though there was some separation, the viscosities remainedquite stable. We did note some shear thinning in the Test samples. Whensamples were allowed to “equilibrate” in a beaker the viscosity washigher versus if the sample was shaken just before the viscositymeasurement. We didn't find this to the same extent with the Controldressing. It was possible to reduce the fat in the model dressing to 10%or “low fat” by increasing the alternan to 6.5%. The resulting dressinghad similar characteristics to the Control with respect to viscosity andappearance. There was some slight separation after being stored at 37.8°C. for 7 days but the viscosity remained quite consistent.

2.3 Bakery Goods

Background:

The objective was to test the alternan as a fat replacer in bakerygoods. Two model systems (cookie and cake) were tested. For cookie thefat replacement was at 30% and for cake at 30% and 50%.

Sugar Snap Cookie

Formula and Methods

Ingredient Full fat (g) % Test (g) % Wheat flour standard 150.0 45.4150.0 45.4 Water 25.5 7.7 35.8 10.8 Soda 1.2 0.4 1.2 0.4 Milk powder 7.52.3 7.5 2.3 Baking powder 2.3 0.7 2.3 0.7 Salt 1.5 0.5 1.5 0.5Shortening 67.5 20.4 47.3 14.3 Sugar 75.0 22.7 75.0 22.7 Alternan — 10.03.0 sum 330.5 100.0 330.5 100.0 Fat % 21.0 15.0 Fat reduction % — 30.0

Method 1:

a) Full fat cookie (control)

-   -   All solids (flour, sugar, soda, baking powder, salt, milk        powder) were mixed in farinograph for 30 sec    -   Shortening added and mixed for 1 min and 30 sec    -   Water added and mixed for 2 min and 30 sec    -   Dough rolled (height 1 cm) and 4 cookies cut with a cutter (76.2        mm)    -   Cookies placed on baking plan and baked at 193° C. for 10 min

b) Alternan reduced fat cookie

-   -   As described in the control except that alternan powder was        mixed with solids

Method 2:

a) Alternan reduced fat cookie

-   -   Making Alternan gel 22% (Alteman powder added to water and        sheared with ULTRA TURRAX® at 11600 rpm for 5 min)    -   All solids (flour, sugar, soda, baking powder, salt, milk        powder) were mixed in farinograph for 30 sec    -   Shortening added and mixed for 1 min and 30 sec    -   Alternan gel added and mixed for 2 min and 30 sec    -   Dough rolled (height 1 cm) and 4 cookies cut with a cutter (76.2        mm)    -   Cookies placed on baking plan and baked at 193° C. for 10 min.

Results and Observations:

Parameter Full Fat Test 1 Test 2 Dough weight g 326.6 326.8 324.9 Bakeddough g 227.6 241.5 242.9 Dough appearance regular regularregular-firmly Cookie weight g 211.7 223.4 224.1 Cookie diameter cm 8.37.7 7.9 Height of all cookies cm 5.0 5.3 5.4 Cookie volume cm³ 270.4246.7 264.6 Baking loss % 7.0 7.5 7.7 Spread ratio 6.6 5.8 5.9 Shaperound slightly oval round Browning normal normal normal Surface smoothcoarse coarse Hardness of cookie fresh kg 3.5 5.6 7.1 week kg 6.1 6.28.6 Crumb moisture fresh % 9.8 16.1 16.4 week % 9.8 12.8 11.3 Test 1 =Cookies with Alternan by method 1 Test 2 = Cookies with Alternan bymethod 2

Cake (Sand Cake)

Formulas and Methods

Full fat Test Test Test Test Test Ingredients % 1% 2% 3% 4% 5% Wheatflour 14 14 14 14 14 14 standard Potato starch 14 14 14 14 14 14Shortening 25 17.5 12.5 12.5 12.5 12.5 Sugar 21 21 21 21 21 21 Vanillasugar 1.4 1.4 1.4 1.4 1.4 1.4 Whole egg 24 24 24 24 24 24 Salt 0.1 0.10.1 0.1 0.1 0.1 Baking powder 0.5 0.5 0.5 0.5 0.5 0.5 Alternan — 1.5 1.53 3 6 Water — 6 11 9.5 9.5 6.5 Sum 100 100 100 100 100 100 Fat % 28 20.515.5 15.5 15.5 15.5 Fat 27 45 45 45 45 reduction %

Method 1:

a) Full fat cake (control)

-   -   The solids (flour, salt, starch and baking powder were blended        (mixture)    -   The shortening, sugar and vanilla sugar were mixed in a kitchen        machine for 1 min    -   The whisked eggs then added and mixed for 1 min    -   The mixture was gradually added and mixed for 2 min    -   120 g of dough was placed in a baking pan and baked for 40 min        at 160° C.

b) Alternan reduced fat cake (Tests 1 to 3 and Test 5)

-   -   As described in the control, except that alternan powder was        mixed with solids

Method 2:

a) Alternan reduced fat cookie (Test 4)

-   -   Making Alternan gel 24% (Alternan powder added to water and        sheared with ULTRA TURRAX® at 11600 rpm for 5 min)    -   The solids (flour, salt, starch and baking powder were blended        (mixture)    -   The Alternan gel, shortening, sugar and vanilla sugar were mixed        in a kitchen machine for 1 min.

Results and Observations:

Parameter Full fat Test 1 Test 2 Test 3 Test 4 Test 5 Dough consistencycreamy/firm less firm less firm less firm less firm less firm Doughmoisture % 21.7 27.7 31.3 30.8 30.4 29 Cake weight g 106.9 104.4 102.7103.2 102.9 103.6 Baking loss % 10.9 13 14.4 14 14.25 13.7 Cake volumeml 575 500 420 420 410 515 Cake moisture % % 12.1 16.9 19.7 19.5 18.817.8 Crumb moisture % fresh 21.1 27.3 33.6 31.4 32.2 28.4 week 12 18.122.5 22.2 23.2 19.1 Cake height mm 50.2 47.6 44.1 46.4 44.9 48.1 Crumbfirmness g fresh 337 296 313 285.4 319.3 213.9 week 1282 1162 1255 11701028 1285 Cake browning golden brown golden brown golden brown goldenbrown golden brown golden brown Crumb appearance succulent/drysucculent/less succulent/less succulent/slightly irregularlysucculent/less (normal) dry dry moist baked dry

The invention claimed is:
 1. A method of emulsifying a product comprising adding an alternan having a weight average molecular weight (Mw) in the range of 10 000 000 g/mol to 60 000 000 g/mol, as determined with GPC MALLS.
 2. The method according to claim 1, wherein the alternan has a weight average molecular weight (Mw) in the range of 33 000 000 g/mol to 60 000 000 g/mol, as determined with GPC MALLS.
 3. The method according to claim 1, wherein the alternan has a weight average molecular weight (Mw) in the range of 12 000 000 g/mol to 30 000 000 g/mol, as determined with GPC MALLS.
 4. The method according to claim 1, wherein the product is a foodstuff.
 5. The method according to claim 4, wherein the foodstuff is a dairy product, yoghurt, ice cream, milk-based soft ice, milk-based garnish, pudding, cream, whipped cream, chocolate cream, butter cream, crème fraiche, curd, milkshakes, egg custard, cheese, nutrition bar, energy bar, breakfast bar, confectionery, bakery product, cracker, cookie, biscuit, cereal chip, snack product, infant and baby foodstuff, bread, croissant, spread, sugar-free biscuit and chocolate, calcium chew, meat product, sausage, mayonnaise, dressing, nut butter, deep-frozen meal, sauce, gravy, soup, shortening, canned food or ready-to-serve meal.
 6. The method of claim 5, wherein the cheese is a cream cheese, soft cheese, sliced cheese, or hard cheese.
 7. The method according claim 1, wherein the product is a cosmetic preparation.
 8. The method according to claim 7, wherein the cosmetic preparation is a deodorant, antiperspirant, antiperspirant/deodorant, shaving product, skin cream and lotion, moisturizer, toner, bath product, cleansing product, hair care product, nail cream and lotion, cuticle softener, protective cream, and other personal care formulations.
 9. The method of claim 8, wherein the hair care product is a shampoo, conditioner, mousse, or styling gel.
 10. The method of claim 8, wherein the protective cream is a sunscreen or anti-aging product. 